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研究一种不使用苯酚的荚膜多糖提制工艺,用于疫苗制备。采用液体罐培养A群和C群脑膜炎球菌,杀菌后离心取上清液,用十六烷基三甲基溴化铵沉淀多糖,用氯化钙溶液解离后,加乙醇至25%去除核酸等杂质,经乙醇沉淀获得粗多糖。将粗多糖溶解后,加乙醇至一定浓度沉淀蛋白,离心获得上清液,经超滤浓缩和乙醇沉淀,获得纯化多糖。纯化的多糖生化检测结果符合WHO和《中国药典》(2010年版)规定,豚鼠和小鼠异常毒性检查合格,豚鼠过敏试验未见过敏反应,1H NMR检测显示多糖结构具有一致性。该研究建立的A群和C群脑膜炎球菌荚膜多糖提制工艺为应用于疫苗制备奠定了基础。
To study a phenol-free capsular polysaccharide preparation process for vaccine preparation. The liquid cans were used to culture group A and group C meningococci. After sterilization, the supernatant was centrifuged, and the polysaccharides were precipitated with cetyltrimethylammonium bromide. After dissociation with calcium chloride solution, ethanol was added to 25% to remove Nucleic acids and other impurities, obtained by ethanol precipitation crude polysaccharides. After the crude polysaccharide is dissolved, ethanol is added to precipitate the protein at a certain concentration, the supernatant is obtained by centrifugation, and the purified polysaccharide is obtained by ultrafiltration concentration and ethanol precipitation. Purified polysaccharide biochemical test results in line with the WHO and the “Chinese Pharmacopoeia” (2010 Edition) provides that guinea pigs and mice were tested for abnormal toxicity, no allergic reactions in guinea pig allergy tests, 1H NMR showed polysaccharide structure is consistent. The study established the group A and group C meningococcal capsular polysaccharide preparation process for the preparation of the vaccine laid the foundation.